Departments of Ophthalmology, Sapporo Medical University School of Medicine, Hokkaido 060-8556, Japan.
Cardiovascular, Renal and Metabolic Medicine, Sapporo Medical University School of Medicine, Hokkaido 060-8556, Japan.
Curr Issues Mol Biol. 2021 Oct 22;43(3):1715-1725. doi: 10.3390/cimb43030121.
In the current study, to elucidate the pathological characteristics of myopic scleral stroma, three-dimensional (3D) cultures of human scleral stroma fibroblasts (HSSFs) with several axial lengths (AL, 22.80-30.63 mm) that were obtained from patients ( = 7) were examined. Among the three groups of ALs, <25 mm ( = 2), 25-30 mm ( = 2), and >30 mm ( = 3), the physical properties of the 3D HSSFs spheroids with respect to size and stiffness, the expressions of extracellular matrix (ECM) molecules, including collagen (COL) 1, 4, and 6 and fibronectin (FN) by qPCR and immunohistochemistry (IHC), and the mRNA expression of ECM metabolism modulators including hypoxia-inducible factor 1A (HIF 1A), HIF 2A, lysyl oxidase (LOX), tissue inhibitor of metalloproteinase (TIMP) 1-4, and matrix metalloproteinase (MMP) 2, 9, and 14 as well as several endoplasmic reticulum (ER) stress-related factors were compared. In the largest AL group (>30 mm), the 3D HSSFs spheroids were (1) significantly down-sized and less stiff compared to the other groups, and (2) significant changes were detected in the expression of some ECMs (qPCR; the up-regulation of and , and the down-regulation of , IHC; the up-regulation of COL1 and FN, and down-regulation of COL4). The mRNA expressions of ECM modulators and ER stress-related genes were also altered with increasing AL length (up-regulation of , and , down-regulation of and , , , and ). In addition, a substantial down-regulation of some ER stress-related genes (, and ) was observed in the 25-30 mm AL group. The findings presented herein suggest that small and stiffer 3D HSSFs spheroids in the largest AL group may accurately replicate the pathological significance of scleral thinning and weakening in myopic eyes. In addition, the modulation of several related factors among the different AL groups may also provide significant insights into our understanding of the molecular mechanisms responsible for causing myopic changes in the sclera.
在本研究中，为了阐明近视性巩膜基质的病理特征，我们培养了来自于患者（=7）的具有不同眼轴长度（AL，22.80-30.63mm）的三维（3D）人巩膜基质成纤维细胞（HSSF），并对其进行了研究。在这三组 AL 中，<25mm（=2）、25-30mm（=2）和>30mm（=3），我们比较了 3D HSSF 球体的大小和硬度、细胞外基质（ECM）分子的表达，包括胶原蛋白（COL）1、4 和 6 以及纤维连接蛋白（FN）的 qPCR 和免疫组织化学（IHC），以及 ECM 代谢调节剂（包括缺氧诱导因子 1A（HIF 1A）、HIF 2A、赖氨酰氧化酶（LOX）、金属蛋白酶组织抑制剂（TIMP）1-4 和基质金属蛋白酶（MMP）2、9 和 14）以及几种内质网（ER）应激相关因子的 mRNA 表达。在最大 AL 组（>30mm）中，3D HSSF 球体（1）与其他组相比明显缩小且硬度降低，（2）一些 ECM 的表达发生了显著变化（qPCR；上调和，下调，IHC；COL1 和 FN 的上调，COL4 的下调）。ECM 调节剂和 ER 应激相关基因的 mRNA 表达也随着 AL 长度的增加而改变（上调、和，下调和、、、和）。此外，在 25-30mmAL 组中，一些 ER 应激相关基因（、和）的表达也显著下调。本研究结果表明，在最大 AL 组中，较小且硬度较大的 3D HSSF 球体可能准确复制了近视眼中巩膜变薄和弱化的病理意义。此外，不同 AL 组之间的几种相关因素的调节也可能为我们理解导致巩膜近视性改变的分子机制提供重要线索。
